In 2019, 4.95 million deaths were associated with antimicrobial resistance (AMR) worldwide. The ESKAPE pathogens, comprised of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. are the primary bacterial contributors. The human microbiome has been identified as a source of acquired resistance in ESKAPE pathogens, in particular the gastrointestinal microbiome, which is host to the highest abundance and diversity of bacteria within the human body. Previously, a high proportion of the bacteria within the gastrointestinal microbiome were unculturable. Consequently, the only method of identifying antimicrobial resistance in gastrointestinal microbiota was through metagenomics. Although metagenomics can identify any known AMR genes, it cannot identify novel AMR genes or determine their expression and functionality within the cell. To overcome these limitations, our study utilised novel culturing on YCFA media in anaerobic conditions, which has been shown to allow for 96% of the gastrointestinal microbiota to be cultured. This method was used to culture 10 faecal samples from healthy individuals, with and without six common orally administered antimicrobials (Amoxicillin, Amoxicillin-Clavulanic acid, Cefalexin, Ciprofloxacin, Clindamycin and Doxycycline). The resultant 1058 colonies were picked, identity determine through 16s rRNA sequencing and resistance to each of the antimicrobials was confirmed in broth. High levels of antimicrobial resistance were determined, with 47.5% of isolates resistant Amoxicillin, 6.3% to Amoxicillin-Clavulanic acid, 44.3% to Cefalexin, 28.8% to Ciprofloxacin, 38.5% to Clindamycin and 25.5% to Doxycycline. Coupled with whole genome sequencing of resistant isolates and comparative genomics, this project aims to identify both known and novel AMR genes, and phenotypically validate this resistance, to demonstrate the diversity of AMR within the human gastrointestinal microbiota. This will allow for identification of the genetic elements responsible for phenotypic AMR within the microbiome, providing a greater understanding for how the microbiome may be spreading AMR.